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Cloning the simian varicella virus genome in E. coli as an infectious bacterial artificial chromosome
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Cloning the simian varicella virus genome in E. coli as an infectious bacterial artificial chromosome
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Journal Article
Cloning the simian varicella virus genome in E. coli as an infectious bacterial artificial chromosome
2011
Overview
Simian varicella virus (SVV) is closely related to human varicella-zoster virus and causes varicella and zoster-like disease in nonhuman primates. In this study, a mini-F replicon was inserted into a SVV cosmid, and infectious SVV was generated by co-transfection of Vero cells with overlapping SVV cosmids. The entire SVV genome, cloned as a bacterial artificial chromosome (BAC), was stably propagated upon serial passage in
E. coli
. Transfection of pSVV-BAC DNA into Vero cells yielded infectious SVV (rSVV-BAC). The mini-F vector sequences flanked by
loxP
sites were removed by co-infection of Vero cells with rSVV-BAC and adenovirus expressing
Cre
-recombinase. Recombinant SVV generated using the SVV-BAC genetic system has similar molecular and
in vitro
replication properties as wild-type SVV. To demonstrate the utility of this approach, a SVV ORF 10 deletion mutant was created using two-step Red-mediated recombination. The results indicate that SVV ORF 10, which encodes a homolog of the HSV-1 virion VP-16 transactivator protein, is not essential for
in vitro
replication but is required for optimal replication in cell culture.
Publisher
Springer Vienna,Springer,Springer Nature B.V
Subject
/ Animals
/ Bacterial artificial chromosomes
/ Biological and medical sciences
/ Biomedical and Life Sciences
/ Cercopithecine alphaherpesvirus 9
/ Chromosomes, Artificial, Bacterial
/ Cosmids
/ DNA
/ E coli
/ Fundamental and applied biological sciences. Psychology
/ genetics
/ genome
/ Genomes
/ humans
/ mutants
/ Primates
/ proteins
/ replicon
/ virion
/ Virions
/ Virology
/ Viruses
